Expression of mRNAs coding for VAP1/crotastatin-like metalloproteases in the venom glands of three South American pit vipers assessed by quantitative real-time PCR.
Snake venom metalloproteases encompass a large family of toxins, with approximately 200 members already catalogued, which exhibit a diversity of structures and biological functions. From this relatively large number, only a dozen examples of apoptosis-inducing metalloproteases, like VAP1 and 2 from the venom of Crotalus atrox, are known. Since most VAP1-like toxins ever characterized were purified from the venom of Viperidae species inhabiting diverse places on earth, we investigate the expression of VAP-like metalloproteases in the venom gland of three representative pit vipers of the Brazilian territory. By molecular cloning and quantitative real-time polymerase chain reaction, using as calibrator gene the Crotalus durissus terrificus homolog of VAP1, named crotastatin, it is reported here that VAP1/crotastatin-like homologues in the venom gland of Bothrops atrox, C. d. cascavella and Lachesis m. rhombeata are expressed at different levels. Hence, batroxstatins, the crotastatin-like precursors from B. atrox, are expressed 87 times more than crotastatin-1, from C. d. cascavella, and 7.5-fold that lachestatins, from L. m. rhombeata. Moreover, in silico structural analysis of amino acid sequences indicates that batroxstatin-2, crotastatins and lachestatin-1 and -2 which share the archetypal motifs and metal- binding sites of VAP1, are subgrouped in a branch that comprises some apoptosis-inducing toxins.
Tavares
N A C
NA
Department of Biochemistry and Laboratory of Immunopathology Keizo Asami - LIKA, Federal University of Pernambuco, Av. Professor Moraes Rego 1235, Cidade Universitária, Recife, PE 50670-901, Brazil.
Correia
J M
JM
Guarnieri
M C
MC
Lima-Filho
J L
JL
Prieto-da-Silva
A R B
AR
Rádis-Baptista
G
G
D000595:Amino Acid Sequence; D000818:Animals; D051017:Apoptosis Regulatory Proteins / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D003198:Computer Simulation; D003435:Crotalid Venoms / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D017839:Crotalus / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D015723:Gene Library; D016014:Linear Models; D008666:Metalloendopeptidases / Q000235:genetics* / Q000378:metabolism; D045726:Metalloproteases / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D012333:RNA, Messenger / Q000032:analysis; D016415:Sequence Alignment
D051017:Apoptosis Regulatory Proteins; D003435:Crotalid Venoms; D012333:RNA, Messenger; D045726:Metalloproteases; D008666:Metalloendopeptidases; C417254:vascular apoptosis-inducing protein 1, Crotalus atrox